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    Cocaine derivative, protein conjugate thereof, monoclonal antibody producing cell line, method for preparing the cell line and monoclonal antibody
    1.
    发明授权
    Cocaine derivative, protein conjugate thereof, monoclonal antibody producing cell line, method for preparing the cell line and monoclonal antibody 失效
    可卡因衍生物，其蛋白质缀合物，产生单克隆抗体的细胞系，制备细胞系的方法和单克隆抗体

    公开(公告)号：US06174723B1

    公开(公告)日：2001-01-16

    申请号：US08982963

    申请日：1997-12-02

    申请人： Keiko Yugawa ,  Nobuyuki Sigetoh ,  Jinsei Miyazaki ,  Tadayasu Mitsumata

    发明人： Keiko Yugawa ,  Nobuyuki Sigetoh ,  Jinsei Miyazaki ,  Tadayasu Mitsumata

    IPC分类号： C12N506

    CPC分类号： G01N33/946 ,  A61K47/60 ,  A61K47/643 ,  C07D451/12 ,  C07K16/16

    摘要： A method for preparing a cocaine-protein conjugate easily by using a cocaine derivative having a methoxy carbonyl group and benzoyl group. This conjugate is useful for the detection of cocaine or cocaine derivatives. A monoclonal antibody, a monoclonal antibody producing cell line, and a method for producing the monoclonal antibody producing cell line by using the above cocaine-protein conjugate as an immunogen is also described. The method of the invention comprises preparing a cocaine-protein conjugate with a pyridyl dithiococaine derivative of Formula (I): where R is either H or R′; and where R′ is: wherein n=1 to 4 and Ph is a benzene ring, and wherein said monoclonal antibody detects 10−9 M cocaine; and which derivative binds to a protein by a disulfide bond; immunizing A/J mouse with the conjugate; fusing its spleen cell and a myeloma cell line; cloning the fusion to produce a monoclonal antibody producing cell line; culturing the cell line, and purifying a supernatant of the culture to form a monoclonal antibody to specifically bind to cocaine or cocaine derivatives.

    摘要翻译： 通过使用具有甲氧基羰基和苯甲酰基的可卡因衍生物容易地制备可卡因 - 蛋白质结合物的方法。 该缀合物可用于检测可卡因或可卡因衍生物。 还描述了单克隆抗体，产生单克隆抗体的细胞系，以及通过使用上述可卡因蛋白质缀合物作为免疫原来制备产生单克隆抗体的细胞系的方法。 本发明的方法包括用式（I）的吡啶基二硫代可卡因衍生物制备可卡因蛋白质结合物：其中R是H或R'; 其中R'为：其中n = 1-4，Ph为苯环，其中所述单克隆抗体检测10-9M可卡因; 哪个衍生物通过二硫键与蛋白质结合; 用缀合物免疫A / J小鼠; 融合其脾细胞和骨髓瘤细胞系; 克隆融合物以产生产生单克隆抗体的细胞系; 培养细胞系，并纯化培养物的上清液以形成特异性结合可卡因或可卡因衍生物的单克隆抗体。